Identification of Formyl Kynurenine Formamidase and Kynurenine Aminotransferase from Saccharomyces cerevisiae Using Crystallographic, Bioinformatic and Biochemical Evidence
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- 作者：Mark Wogulis ; Erin R. Chew ; Paul D. Donohoue ; David K. Wilson
- 刊名：Biochemistry
- 出版年：2008
- 出版时间：February 12, 2008
- 年：2008
- 卷：47
- 期：6
- 页码：1608 - 1621
- 全文大小：1215K
- 年卷期：v.47,no.6(February 12, 2008)
- ISSN：1520-4995
文摘
The essential enzymatic cofactor NAD+ can be synthesized in many eukaryotes, includingSaccharomyces cerevisiae and mammals, using tryptophan as a starting material. Metabolites along thepathway or on branches have important biological functions. For example, kynurenic acid can act as anNMDA antagonist, thereby functioning as a neuroprotectant in a wide range of pathological states. N-Formylkynurenine formamidase (FKF) catalyzes the second step of the NAD+ biosynthetic pathway by hydrolyzingN-formyl kynurenine to produce kynurenine and formate. The S. cerevisiae FKF had been reported to bea pyridoxal phosphate-dependent enzyme encoded by BNA3. We used combined crystallographic,bioinformatic and biochemical methods to demonstrate that Bna3p is not an FKF but rather is most likelythe yeast kynurenine aminotransferase, which converts kynurenine to kynurenic acid. Additionally, weidentify YDR428C, a yeast ORF coding for an 
hars/alpha.gif" BORDER=0>/hars/beta2.gif" BORDER=0 ALIGN="middle"> hydrolase with no previously assigned function, asthe FKF. We predicted its function based on our interpretation of prior structural genomics results and onits sequence homology to known FKFs. Biochemical, bioinformatics, genetic and in vivo metabolite dataderived from LC-MS demonstrate that YDR428C, which we have designated BNA7, is the yeast FKF.
hars/alpha.gif" BORDER=0>/hars/beta2.gif" BORDER=0 ALIGN="middle"> hydrolase with no previously assigned function, asthe FKF. We predicted its function based on our interpretation of prior structural genomics results and onits sequence homology to known FKFs. Biochemical, bioinformatics, genetic and in vivo metabolite dataderived from LC-MS demonstrate that YDR428C, which we have designated BNA7, is the yeast FKF.